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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.

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Related Experiment Video

Updated: Jun 6, 2026

Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy
13:13

Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy

Published on: December 3, 2012

Protein localization in electron micrographs using fluorescence nanoscopy.

Shigeki Watanabe1, Annedore Punge, Gunther Hollopeter

  • 1Department of Biology and Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah, USA.

Nature Methods
|November 25, 2010
PubMed
Summary
This summary is machine-generated.

Researchers developed a new method combining fluorescence and electron microscopy to precisely map protein locations within cells. This technique overcomes limitations of traditional methods, enabling detailed molecular topography for better cell understanding.

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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

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The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells
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The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

Published on: February 27, 2020

Related Experiment Videos

Last Updated: Jun 6, 2026

Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy
13:13

Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy

Published on: December 3, 2012

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells
11:45

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

Published on: February 27, 2020

Area of Science:

  • Cell Biology
  • Microscopy
  • Molecular Biology

Background:

  • Understanding cellular molecular topography requires linking proteins to specific organelles.
  • Immunocytochemical electron microscopy offers nanometer resolution but faces limitations in fixation quality, antibody availability, and antigen accessibility.

Purpose of the Study:

  • To develop and demonstrate correlative fluorescence electron microscopy for nanoscopic protein localization in electron micrographs.
  • To overcome limitations of existing protein localization techniques.

Main Methods:

  • Proteins were tagged with fluorescent proteins (Citrine or tdEos) and expressed in Caenorhabditis elegans.
  • Fixed worms were embedded in plastic, sectioned, and imaged using stimulated emission depletion (STED) microscopy or photoactivated localization microscopy (PALM).
  • Fluorescence signals were correlated with organelles visualized in electron micrographs from identical sections.

Main Results:

  • Demonstrated successful nanoscopic localization of tagged proteins within cellular structures.
  • Correlated fluorescence imaging with electron microscopy on ultrathin sections.
  • Successfully localized histones, a mitochondrial protein, and a presynaptic dense projection protein.

Conclusions:

  • Correlative fluorescence electron microscopy provides a powerful approach for detailed molecular mapping within cells.
  • This method enhances the understanding of protein distribution and cellular architecture.
  • The technique offers improved resolution and overcomes previous limitations in protein localization studies.